RNA

Part:BBa_K3380509:Design

Designed by: Alexandru Popov   Group: iGEM20_Edinburgh   (2020-10-16)


No scaffold iSpinach aptamer under T7RNAP promoter (BBa_z0251) and T7 terminator (BBa_K3137010)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 25


Design Notes

This construct has not been tested in the lab, however the Edinburgh iGEM 2020 team tested the BBa_K3380500 construct. The BBa_K3380509 construct can be assembled following a similar protocol (on the experience page of the BBA_K3380500 construct).

All the sequences for the individual parts were acquired. The individual part were designed with 4bp scars to allow swapping of the parts to test different constructs. The individual parts were phosphorylated, annealed and ligated together using the T4 ligase.

Source

The parts were acquired as de novo synthesized sequences from Sigma-Aldrich.

References

Millacura, F.A., Li, M., Valenzuela-Ortega, M.A. and French, C., 2019. TXO: Transcription-Only genetic circuits as a novel cell-free approach for Synthetic Biology. bioRxiv, p.826230.

Autour, A., Westhof, E. and Ryckelynck, M., 2016. iSpinach: a fluorogenic RNA aptamer optimized for in vitro applications. Nucleic acids research, 44(6), pp.2491-2500